Establishment of an improved model of oxygen glucose deprivation/reoxygenation of primary hippocampal neurons
Objective To establish an improved model of oxygen glucose deprivation/reoxygenation(OGD/R)in primary hippocampal neurons.Methods Hippocampal neurons were isolated from brain tissue ofSD neonatal rat,and neuron suspension was prepared.By inhibiting the number of glial cells,the neuron suspension was divided into three groups,and no inhibition,partial inhibition and complete inhibition,the cell morphology was observed under microscope to determine the optimal culture conditions for primary hippocampal neurons.Based on this condition,the hippocampal neurons were deprived of glucose and oxygen for 0.5,1 and 1.5 h,and then reoxygenated for 0,24,36 and 48 h,separately.The content of lactic dehydrogenase(LDH)in culture medium was detected by ELISA,and the survival rate of neurons was measured by CCK-8 assay.The condition with higher LDH content and less influence on survival rate was the optimal condition for OGD/R model of primary hippocampal neurons.Results When the glial cells were partially inhibited,the purity of neurons was high with uniform growth status.The ratio of glial cells to hippocampal neurons was about 1∶1,which preserved the structural and functional connection between glial cells and neurons.Compared with OGD 1 h group,the LDH content increased signifi-cantly at OGD 1 h/R36h(t=4.31,P＜0.05),while there was no significant difference in neuronal survival rate(t=1.99,P＞0.05).Conclusion Primary hippocampal neurons with about 1∶1 number of glial cells and neurons were obtained with good growth status.The OGD/R model established under the condition of OGD 1 h/R 36 h has relatively high survival rate with shortened dendrites and impaired vitality,which lays a foundation for the further study of the pathogenesis of related diseases.
NETL
NSTL
万方数据
